An Investigation into the Methods of Detection of Toxocara Species Eggs in the Environment and the Differentiation of G217B and WU24 Strains of Histoplasma Capsulatum by Restriction Fragment Length Polymorphisms in the CBP Gene

Abstract

Toxocariasis is a human parasitic disease caused by infection with the larva of Toxocara canis and Toxocara cati with adult worms found in dogs and cats, respectively. With few consistent signs or symptoms, there is an underestimate of the human population that is affected by this disease (Bellanger et al., 2009; Gordon et al., 2011). The level of environmental contamination is directly related to the risk of human infection (Misgajska, 2001), making testing for this contamination imperative to understanding the prevalence of this disease. The original goals of this experiment were to obtain sand samples from parks and playgrounds in Cleveland to determine if Toxocara species eggs were present through DNA amplification. Macuhova et al. designed a novel, more sensitive, cheaper and faster method to test for T. cati and T.canis eggs using LAMP amplification rather than traditional PCR methods. In the experiments outlined in this thesis, this method failed to produce usable results due to false positive readings. PCR was also used to try to identify Toxocara egg DNA from contaminated fecal samples, but due to a failure in DNA extraction or amplification, these methods were unable to amplify positive control samples obtained from infected animal feces. Future work should include finding a consistent and effective way to extract DNA from environmental samples and amplify the DNA to identify contaminated samples.